VALIDATION IMAGES
Sample:
Lane 1: Human HT-1080 cell lysates
Lane 2: Human HeLa cell lysates
Lane 3: Human JAR cell lysates
Primary: Anti-LAMP1 (bs-1970R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: kDa
Observed band size: 95 kDa
Paraformaldehyde-fixed, paraffin embedded Human Small Intestine; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Liver Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:100 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LAMP1 Polyclonal Antibody, Unconjugated (bs-1970R) at 1:100 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
4% Paraformaldehyde-fixed MCF-7 (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (LAMP1) polyclonal Antibody, unconjugated (bs-1970R) 1:200, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-0295G-FITC) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
Tissue/cell: A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (LAMP1) Polyclonal Antibody, Unconjugated (bs-1970R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-cy3) at 37°C for 90 minutes, DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.
Blank control:Jurkat.
Primary Antibody (green line): Rabbit Anti-LAMP1 antibody (bs-1970R)
Dilution: 2ug/Test;
Secondary Antibody (white blue line) : Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Isotype control(orange line):Normal Rabbit IgG
Protocol
The cells were incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.